Modulated release particles for lung delivery

ABSTRACT

A modulated release aerosol formulation is disclosed. The formulation comprises a polysaccharide polymer having a selected medicament associated therewith, a fluid carrier for carrying and delivering the construct and a stabilizer.

BACKGROUND OF THE INVENTION

[0001] 1 . Field of the Invention

[0002] This invention relates to modulated release aerosol particles,and more particularly, to medicinal aerosol particles comprisingpolymeric vesicles which entrap a selected medicament and provide slowrelease thereof.

[0003] 2 . Description of the Related Art

[0004] Many drugs currently administered by inhalation come primarily asliquid or solid aerosol particles of respirable size. For biotherapeuticdrugs, this may present a problem, as many of these medicaments areunstable in aqueous environments for extended periods of time and arerapidly denatured if micronized by high shear grinding or othercomminution methods when presented as dry powders. Additionally, anumber of these medicaments do not survive long enough in the lung asthey are extracted quickly from the lung environment after they areadministered as inhalation aerosols. Significant drug loss could alsooccur by deactivation either as a result of reactivity of the medicamentwith device and container surfaces, or during aerosolization,particularly in high shear, energy intensive, nebulized systems[Mumenthaler, M, et al., Pharm. Res., 11:12-20 (1994)].

[0005] To overcome these instability problems, many drug and excipientsystems contain biodegradable carriers, such aspoly(lactide-co-glycolides,have been developed for biotherapeuticproteins and peptides [Liu, R., et al., Biotechnol. Bioeng., 37:1 77-184(1991)]. These medicaments, presumably, are adequately protected intheir carrier systems, and thus do not undergo as much denaturation asrealized in aqueous media. Importantly, these polymers prolong drugrelease at the site of absorption so that the effect of the drug is alsosubsequently sustained in the body.

[0006] Most therapeutic peptides and proteins are poorly absorbedthrough biologic membranes even upon formulation with penetrationenhancers, possibly due to a combination of several factors, includinglarge molecular size (i.e., ≧1000 daltons), ionization, high surfacecharge, enzymatic and chemical instability, and low permeability ofabsorption barriers in the body of a patient, e.g. human being or otheranimal. In numerous therapies, drug dosimetry is increased by orders ofmagnitude to achieve minimum systemic concentrations required forefficacy. In other cases the drug product is formulated with exoticabsorption promoters in order to improve permeability across theabsorption barrier. But such formulations usually present serioustoxicological liabilities. The clinical and pharmaceutical chemistrysciences, in an attempt to accomplish the highest level of therapeuticbenefit for these compounds, have resorted to chemical modifications asa principal mode for improving biological activity of these drugs in thebody of the patient. The mode of drug administration to the body hasalso gradually expanded from oral and parenteral to transdermal, rectaland the pulmonary routes of administration, i.e., nose and lung. Successand achievement with these drug delivery approaches are mixed largelydue to lack of acceptance of the newer, complex molecules that must beused for treating difficult diseases of the body, e.g., infections,malignancies, cardiovascular, endocrine, neurologic diseases, and avariety of immunologically compromised diseases, like AIDS.

[0007] Accordingly, what is desired and needed is a fluid propelledformulation system comprising an active pharmaceutical ingredient(“API”) that is stable and protected by a rate-limiting carrier, easilymanufactured, and therapeutically effective when administered as fluiddispersed particles to the lung of a patient, e.g. a human being oranother animal.

SUMMARY OF THE INVENTION

[0008] This invention relates to modulated release aerosol particles,and more particularly, to medicinal, respirable aerosol particlescomprising polysaccharide vesicles which are associated with, e.g. forma part of a construct with or entrap therewithin a selected medicamentand provide slow release thereof.

BRIEF DESCRIPTION OF THE DRAWING

[0009] The nature of the invention will appear more fully from thefollowing detailed description taken in conjunction with the appendeddrawing, in which:

[0010]FIG. 1 is a graphical representation of a plasma glucose profileof a stabilized alginate formulation of recombinant human insulin in NewZealand rabbits; and

[0011]FIG. 2 is a graphical representation of a plasma concentrationprofile of hr-insulin following intravenous and aerosol administrationaccording to the invention.

DETAILED DESCRIPTION OF THE INVENTION

[0012] This application makes reference to U.S. applications Ser. No.09/158,369, filed on Sep. 22, 1998, now U.S. Pat. Nos. 6,136,294 and09/209,228 filed on Dec. 10, 1998, which are incorporated hereinto byreference in their entirety.

[0013] This invention involves stable, modulated release, respirable,aerosolizable particles suitable for delivery of medicaments to thelung, which comprise (1) a medicament or drug, (2) a naturally occurringpolysaccharide polymeric construct into which the drug is associated,i.e. is encapsulated therewithin or being part of the construct, (3) asuitable fluid or propellant, (4) a suitable stabilizer. Thepolysaccharide polymeric construct, modulates release of theencapsulated drug to the body of a patient, e.g. a human being oranother animal, when the formulation is administered to the patient'srespiratory tract.

[0014] A suitable macromolecular medicament or drug is one which issuitable for administration by inhalation, the inhalation being used fororal and nasal inhalation therapy. A stable, colloidal dispersion of amedicament in a fluid, e.g. air, hydrocarbon gases, chlorofluorocarbon(CFC) propellants or non-CFC propellants, such as tetrafluoroethane(HFA-134a) and heptafluoropropane (HFA-227), is described.

[0015] A stabilizer of a polyionic species, such as an amino acid and asmall molecule peptide, as an inactive formulation component, whichtriggers loss of adhesive bond strength between the medicamentparticles, may optionally be employed. An electret or steriallystabilized aerocolloid particles of the selected medicaments is thusformed. An electret is the electrostatic equivalent of a permanentmagnet but can be susceptible to breakdown in the presence of moisture,such as that present in air or at ambient humidity conditions of therespiratory tract. Accordingly the present invention applies toparticles formulated for use in dry powder aerosols, portable nebulizersystems, as well pressurized metered dose inhaler formulations.

[0016] The resultant aerocolloid is chemically and physically stable andcan remain in suspension until the selected medicament or drug particlesreach the alveolar or other absorption sites in the airways of apatient, e.g. human, other animal, being treated. Once at the absorptionsite, the drug particles should be efficiently trapped at the depositionsite as a result of moisture in the ambient, dissolve rapidly in theepithelial lining fluids, and be absorbed quickly across thebiomembranes of the patient, thereby limiting possible deactivation bymetabolizing enzymes in the airways.

[0017] As used herein the following terms are defined as follows.

[0018] The term “rate of release” from the polysaccharide polymermedicament carrier is defined as the amount of medicament released perunit time either to the lung environment or from the lung environment tothe systemic circulation of the body of the patient treated.

[0019] The terms “peptide”, “polypeptide”, “oligopeptide” and “protein”shall be used interchangeably when referring to peptide or protein drugsand shall not be limited as to any particular molecular weight, peptidesequence or length, field of bioactivity or therapeutic use unlessspecifically stated.

[0020] A suitable medicament to which the subject invention is directedincludes a peptide, polypeptide, or protein biotherapeutic medicamentranging from 0.5 K Dalton to 150 K Dalton in molecular size. Inparticular, the peptide, polypeptide, or protein biotherapeuticmedicament includes diabetic aids; such as insulins and insulin analogs;amylin; glucagon; surfactants; immunomodulating peptides such ascytokines, chemokines, lymphokines; interleukins, such as taxol,interleukin-1, interleukin-2, and interferons; erythropoetins;thrombolytics and heparins; anti-proteases, antitrypsins and amiloride;rhDNase; antibiotics and other antiinfectives; hormones; and growthfactors, such as parathyroid hormones, LH-RH and GnRH analogs; nucleicacids; DDAVP; calcitonins; cyclosporine; ribavirin; enzymes; heparins;hematopoietic factors; cyclosporins; vaccines; immunoglobulins;vasoactive peptides; antisense agents; genes, oligonucleotide, andnucleotide analogs.

[0021] The term “diabetic aid” includes natural, synthetic,semi-synthetic and recombinant medicaments such as activin, glucagon,insulin, somatostatin, proinsulin, amylin, and the like.

[0022] The term “insulin” shall be interpreted to encompass insulinanalogs, natural extracted human insulin, recombinantly produced humaninsulin, insulin extracted from bovine and/or porcine sources,recombinantly produced porcine and bovine insulin and mixtures of any ofthese insulin products. The term is intended to encompass thepolypeptide normally used in the treatment of diabetics in asubstantially purified form but encompasses the use of the term in itscommercially available pharmaceutical form, which includes additionalexcipients. The insulin is preferably recombinantly produced and may bedehydrated (completely dried) or in solution.

[0023] The terms “insulin analog”, “monomeric insulin” and the like areused interchangeably herein and are intended to encompass any form of“insulin” as defined above, wherein one or more of the amino acidswithin the polypeptide chain has been replaced with an alternative aminoacid and/or wherein one or more of the amino acids has been deleted orwherein one or more additional amino acids has been added to thepolypeptide chain or amino acid sequences, which act as insulin indecreasing blood glucose levels. In general, the term “insulin analogs”of the present invention include “insulin lispro analogs”, as disclosedin U.S. Pat. No. 5,547,929, incorporated hereinto by reference in itsentirety; insulin analogs including LysPro insulin and humalog insulin,and other “super insulin analogs”, wherein the ability of the insulinanalog to affect serum glucose levels is substantially enhanced ascompared with conventional insulin as well as hepatoselective insulinanalogs which are more active in the liver than in adipose tissue.Preferred analogs are monomeric insulin analogs, which are insulin-likecompounds used for the same general purpose as insulin, such as insulinlispro, i.e., compounds which are administered to reduce blood glucoselevels.

[0024] The term “amylin” includes natural human amylin, bovine, porcine,rat, rabbit amylin, as well as synthetic, semi-synthetic or recombinantamylin or amylin analogs including pramlintide and other amylinagonists, as disclosed in U.S. Pat. Nos. 5,686,411 and 5,854,215, bothof which are incorporated hereinto by reference in their entirety.

[0025] The term “immunomodulating proteins” include cytokines,chemokines, lymphokines complement components, immune system accessoryand adhesion molecules and their receptors of human or non-human animalspecificity. Useful examples include GM-CSF, IL-2, IL-12, OX40, OX40L(gp34), lymphotactin, CD40, CD40L. Useful examples include interleukins,for example interleukins 1 to 15; interferons alpha, beta or gamma;tumour necrosis factor, granulocyte-macrophage colony stimulating factor(GM-CSF), macrophage colony stimulating factor (M-CSF), granulocytecolony stimulating factor (G-CSF), chemokines, such as neutrophilactivating protein (NAP); macrophage chemoattractant and activatingfactor (MCAF), RANTES, macrophage inflammatory peptides MIP-1a andMIP-1b, complement components and their receptors, or an accessorymolecule, such as B7.1, B7.2, ICAM-1, 2 or 3 and cytokine receptors.OX40 and OX40-ligand (gp34) are further useful examples ofimmunomodulatory proteins. Immunomodulatory proteins can for variouspurposes be of human or non-human animal specificity and can berepresented, for present purposes, as the case may be and as may beconvenient, by extracellular domains and other fragments with thebinding activity of the naturally occurring proteins, and muteinsthereof, and their fusion proteins with other polypeptide sequences,e.g. with immunoglobulin heavy chain constant domains. Where nucleotidesequences encoding more than one immunomodulating protein are inserted,they can, for example, comprise more than one cytokine or a combinationof cytokines and accessory/adhesion molecules.

[0026] The term “interferon” or “IFN” as used herein means the family ofhighly homologous species-specific proteins that inhibit viralreplication and cellular proliferation and modulate immune response.Interferons are grouped into three classes based on their cellularorigin and antigenicity, namely, alpha-interferon (leukocytes),beta-interferon (fibroblasts) and gamma-interferon (immunocompetentcells). Recombinant forms and analogs of each group have been developedand are commercially available. Subtypes in each group are based onantigenic/structural characteristics. At least 24 interferon alphas(grouped into subtypes A through H) having distinct amino acid sequenceshave been identified by isolating and sequencing DNA encoding thesepeptides. Reference is made to Viscomi, 1996 Biotherapy 10:59-86, thecontents of which are incorporated by reference hereinto in itsentirety. The terms “alpha.-interferon”, “alpha interferon”, “interferonalpha”, “human leukocyte interferon” and “IFN” are used interchangeablyherein to describe members of this group. Both naturally occurring andrecombinant alpha interferons, including consensus interferon such asthat described in U.S. Pat. No. 4,897,471, the contents of which areincorporated hereinto by reference in its entirety, may be used in thepractice of the invention. Human leukocyte interferon prepared in thismanner contains a mixture of human leukocyte interferons havingdifferent amino acid sequences. Purified natural human alpha inteferonsand mixtures thereof which may be used in the practice of the inventioninclude but are not limited to Sumiferon RTM interferon alpha-n1available from Sumitomo, Japan; Welfferong interferon alpha-n1 (Ins)available from Glaxo-Wellcome Ltd., London, Great Britain; and AlferonRTM interferon alpha-n3 available from the Purdue Frederick Co.,Norwalk, Conn.

[0027] The term “erythropoietin” applies to synthetic, semi-synthetic,recombinant, natural, human, monkey, or other animal or microbiologicalisolated polypeptide products having part or all of the primarystructural conformation (i.e., continuous sequence of amino acidresidues) and one or more of the biological properties (e.g.,immunological properties and in vivo and in vitro biological activity)of naturally-occurring erythropoietin, including allelic variantsthereof. These polypeptides are also uniquely characterized by being theproduct of procaryotic or eucaryotic host expression (e.g., bybacterial, yeast and mammalian cells in culture) of exogenous DNAsequences obtained by genomic or cDNA cloning or by gene synthesis.Products of microbial expression in vertebrate (e.g., mammalian andavian) cells may be further characterized by freedom from associationwith human proteins or other contaminants which may be associated witherythropoietin in its natural mammalian cellular environment or inextracellular fluids such as plasma or urine. The products of typicalyeast (e.g., Saccaromyces cerevisiae) or procaryote (e.g., E. coli) hostcells are free of association with any mammalian proteins. Dependingupon the host employed, polypeptides of the invention may beglycosylated with mammalian or other eucaryotic carbohydrates or may benonglycosylated. Polypeptides of the invention may also include aninitial methionine amino acid residue (at position −1). Novelglycoprotein products of the invention include those having a primarystructural conformation such as growth hormone, thyroid hormone, thyroidreleasing hormone (TRH), gonadotropin-releasing hormone (GnRH),leuteininzing hormone, leuteininzing hormone-releasing hormone (LHRH,including the superagonists and antagonists, such as leuprolide,deltirelix, gosorelin, nafarelin, danazol, etc.) sourced from natural,human, porcine, bovine, ovine, synthetic, semi-synthetic, or recombinantsources. These also include somatostatin analogs such as octreotide(Sandostatin). Other agents in this category of biotherapeutics includemedicaments for uterine contraction (e.g., oxytocin), diuresis (e.g.,vasopressin), neutropenia (e.g., GCSF), medicaments for respiratorydisorders (e.g., superoxide dismutase), RDS (e.g., surfactants,optionally including apoproteins), and the like.

[0028] The term “enzymes” include recombinant deoxyribonuclease such asDNAse (Genentech) proteases (e.g., serine proteases such as trypsin andthrombin), polymerases (e.g., RNA polymerases, DNA polymerases), reversetranscriptases and kinases, enzymes implicated in arthritis,osteoporosis, inflammatory diseases, diabetes, allergies, organtransplant rejection, oncogene activation (e.g., dihydrofolatereductase), signal transduction, self-cycle regulation, transcription,DNA replication and repair.

[0029] The term “nucleic acids” includes any segment of DNA or RNAcontaining natural or non-naturally occurring nucleosides, or otherproteinoid agents capable of specifically binding to other nucleic acidsor oligonucleotides via complementary hydrogen-bonding and also arecapable of binding to non-nucleic acid ligates. In this regard,reference is made to Bock, L., et al., Nature 355:564-566 (1992) whichreports inhibition of the thrombin-catalyzed conversion of fibrinogen tofibrin using aptamer DNA.

[0030] Examples of biological molecules for which lead molecules can besynthesized sufficiently duplicative of that of a naturally-occurring(e.g., human) erythropoietin to allow possession of one or more of thebiological properties thereof and having an average carbohydratecomposition which differs from that of naturally-occurring (e.g., human)erythropoietin.

[0031] The terms “heparins” and “thrombolytics” include anti-clottingfactors such as heparin, low molecular weight heparin, tissueplasminogen activator (TPA), urokinase (Abbokinase) and other factorsused to control clots.

[0032] The terms “anti-proteases” and “protease-inhibitors” are usedinterchangeably and apply to synthetic, semi-synthetic, recombinant,naturally-occurring or non-naturally occurring, soluble or immobilizedagents reactive with receptors, or act as antibodies, enzymes or nucleicacids. These include receptors which modulate a humoral immune response,receptors which modulate a cellular immune response (e.g., T-cellreceptors) and receptors which modulate a neurological response (e.g.,glutamate receptor, glycine receptor, gamma-amino butyric acid (GABA)receptor). These include the cytokine receptors (implicated inarthritis, septic shock, transplant rejection, autoimmune disease andinflammatory diseases), the major histocompatibility (MHC) Class I andII receptors associated with presenting antigen to cytotoxic T-cellreceptors and/or T-helper cell receptors (implicated in autoimmunediseases) and the thrombin receptor (implicated in coagulation,cardiovascular disease). Also included are antibodies which recognizeself-antigens, such as those antibodies implicated in autoimmunedisorders and antibodies which recognize viral (e.g., HIV, herpessimplex virus) and/or microbial antigens.

[0033] The terms “hormones” and “growth factors” include hormonereleasing hormones such as growth hormone, thyroid hormone, thyroidreleasing hormone (TRH), gonadotropin- releasing hormone (GnRH),leuteininzing hormone, leuteininzing hormone-releasing hormone (LHRH,including the superagonists and antagonists, such as leuprolide,deltirelix, gosorelin, nafarelin, danazol, etc.) sourced from natural,human, porcine, bovine, ovine, synthetic, semi-synthetic or recombinantsources. These also include somatostatin analogs such as octreotide(Sandostatin). Other agents in this category of biotherapeutics includemedicaments for uterine contraction (e.g., oxytocin), diuresis (e.g.,vasopressin), neutropenia (e.g., GCSF), medicaments for respiratorydisorders (e.g., superoxide dismutase), RDS (e.g., surfactants,optionally including apoproteins), and the like.

[0034] The term “enzymes” include recombinant deoxyribonuclease such asDNAse (Genentech) proteases (e.g., serine proteases such as trypsin andthrombin), polymerases (e.g., RNA polymerases, DNA polymerases), reversetranscriptases and kinases, enzymes implicated in arthritis,osteoporosis, inflammatory diseases, diabetes, allergies, organtransplant rejection, oncogene activation (e.g., dihydrofolatereductase), signal transduction, self-cycle regulation, transcription,DNA replication and repair.

[0035] The term “nucleic acids” includes any segment of DNA or RNAcontaining natural or non-naturally occurring nucleosides, or otherproteinoid agents capable of specifically binding to other nucleic acidsor oligonucleotides via complementary hydrogen-bonding and also arecapable of binding to non-nucleic acid ligates. In this regard,reference is made to Bock, L., et al., Nature 355:564-566 (1992) whichreports inhibition of the thrombin-catalyzed conversion of fibrinogen tofibrin using aptamer DNA.

[0036] Examples of biological molecules for which lead molecules can besynthesized and selected and combined in accordance with the inventioninclude, but are not limited to, agonists and antagonists for cellmembrane receptors, neurotransmitters, toxins and venoms, viralepitopes, hormones, opiates, steroids, peptides, enzyme substrates andinhibitors, cofactors, drugs, lectins, sugars, oligonucleotides, nucleicacids, oligosaccharides, lipids, proteins, and analogs of any of theforegoing molecules.

[0037] The term “analog” refers to a molecule, which shares a commonfunctional activity with the molecule to which it is deemed to becomparable and typically shares common structural features as well.

[0038] The term “recombinant” refers to any type of clonedbiotherapeutic expressed in procaryotic cells or a geneticallyengineered molecule, or combinatorial library of molecules which may befurther processed into another state to form a second combinatoriallibrary, especially molecules that contain protecting groups whichenhance the physicochemical, pharmacological, and clinical safety of thebiotherapeutic agent.

[0039] The term “vaccines” refers to therapeutic compositions forstimulating humoral and cellular immune responses, either isolated, orthrough an antigen presenting cell, such as an activated dendritic cell,that is able to activate T-cells to produce a multivalent cellularimmune response against a selected antigen. The potent antigenpresenting cell is stimulated by exposing the cell in vitro to apolypeptide complex. The polypeptide complex may comprise a dendriticcell-binding protein and a polypeptide antigen, but preferably, thepolypeptide antigen is either a tissue-specific tumor antigen or anoncogene gene product. However, it is appreciated that other antigens,such as viral antigens can be used in such combination to produceimmunostimulatory responses. In another preferred embodiment, thedendritic cell-binding protein that forms part of the immunostimulatorypolypeptide complex is GM-CSF. In a further preferred embodiment, thepolypeptide antigen that forms part of the complex is the tumor-specificantigen prostatic acid phosphatase. In still other preferredembodiments, the polypeptide antigen may be any one of the oncogeneproduct peptide antigens. The polypeptide complex may also contain,between the dendritic cell-binding protein and the polypeptide antigen,a linker peptide. The polypeptide complex may comprise a dendriticcell-binding protein covalently linked to a polypeptide antigen, suchpolypeptide complex being preferably formed from a dendritic cellbinding protein, preferably GM-CSF, and a polypeptide antigen. Thepolypeptide antigen is preferably a tissue-specific tumor antigen suchas prostatic acid phosphatase (PAP), or an oncogene product, such asHer2, p22RAS, and p53; however, other embodiments, such as viralantigens, are also within the scope of the invention.

[0040] The term “mmunoglobulins” encompasses polypeptideoligonucleotides involved in host defense mechanisms, such as coding andencoding by one or more gene vectors, conjugating various bindingmoieties of nucleic acids in host defense cells, or coupling expressedvectors to aid in the treatment of a human or animal subject. Themedicaments included in this class of polypeptides include IgG, IgE,IgM, IgD, either individually or in a combination with one another.

[0041] For purposes of the formulations of this invention, which areintended for inhalation into the lungs, the biotherapeutic medicament isassociated with the naturally occurring polysaccharide polymer to whichit is destined to be combined. By “associate” or “associated” is meantthat the medicament is present as a matrix or a part of a polymericconstruct along with the polysaccharide polymer or is encapsulated as amicrosphere in a polysaccharide polymer or in polysaccharide polymericconstruct particle, or is on a surface of such particle, whereby atherapeutically effective amount or fraction (e.g., 95% percent or more)of the biotherapeutic is particulate. Typically, the construct particleshave a diameter of less than about 10 microns, and preferably less thanabout 5 microns, in order that the particles can be inhaled into therespiratory tract and/or lungs of the patient being treated, e.g. ahuman or other animal.

[0042] A suitable polymeric construct is selected. Such a construct isone which will incorporate therein or encapsulate the selectedmedicament, e.g. insulin, amylin, octreotide, erythopoietin,immunoglobulin, leuprolide, glucagon and provide a controlled ormodulated release of the medicament therefrom to the sites of action orapplication of the patient's body, e.g. from the lung to the localsurrounding environment of the human being or other animal.

[0043] A suitable polysaccharide is a polymer selected from the group ofan alginate salt, e.g. Li⁺, Na⁺, K⁺, Ca⁺⁺, NH⁺⁺⁺, NH⁺⁺⁺⁺ etc., such assodium alginate, calcium alginate, sodium-calcium alginate, ammoniumalginate, sodium-ammonium alginate, or calcium-ammonium alginate. Apreferred aliginate modulating releasing agent is ammonium calciumalginate. These materials are typically used in injectable implants andmicrosphere preparations for controlled release. A commercial form ofammonium calcium alginate is Keltose, manufactured and distributed byISP (International Specialty Products, 1361 Alps Road, Wayne, N.J.07470). As used herein, “alginate” means alginic acid, or any of itssalts; or other naturally occuring polysaccharide or carbohydrate basedpolymers such as gum arabic, pectin, galacturonic acid, gum karaya; gumBenjamin, plantago ovata gum; agar; carrageenan; cellulose; gelatin; ora mixture of any of the foregoing polymers.

[0044] Alginates are pharmaceutical excipients generally regarded assafe and used therefore to prepare a variety of pharmaceutical systemswell documented in the patent literature [S. Bloor, U.S. Pat. No.6,166,084; Ikeda, et al., U.S. Pat. No. 6,166,043; Ikeda, et al., U.S.Pat. No. 6,166,042; Fassler, et al., U.S. Pat. No. 6,166,004; Itakura,et al., U.S. Pat. No. 6,165,615;].

[0045] Alginates are naturally occurring polymers comprisingpolysaccharide chains. These polymers have the propensity to absorbwater thus swelling to become gel-like structures in solution. Uponinhalation of the resultant core formulation by a patient being treated,the gel dissolves in the body of such patient, thus releasing its drugpayloads in a dissolution controlled manner. Such a polymer system formsa construct or a matrix when formed in situ with the selected medicamentor medicaments whereby such medicament or medicaments forms part of thematrix or is encapsulated within the matrix. Upon such formation orencapsulation, the medicament, e.g. entrapped insulin, is time-releasedor modulated from the site of action in the body, e.g. the lungs, therespiratory tract, node, ear, etc., to the surrounding environment ortissues of the body of the patient treated.

[0046] The polysaccharide polymer, e.g. an alginate salt, is typicallypresent in the resultant controlled-release formulation in an amountranging from about 0.000001% to about 10% by weight of the total weightof the formulation.

[0047] The biotherapeutic medicament is present in the inventive polymerconstruct in a therapeutically effective amount, that is, an amount suchthat the biotherapeutic medicament can be incorporated into an aerosolformulation such as a dispersion aerosol, via oral or nasal inhalation,and cause its desired therapeutic effect, typically preferred with onedose, or through several doses.

[0048] The term “dosing period” shall be interpreted to mean the periodduring which administration of the selected medicament may be given to apatient in need thereof by the intrapulmonary route of administrationwhich period may encompass preferably one or more hours in a day or afew days to several weeks but less preferably over a month or under 1hour, but during which period multiple inhalations are made by thepatient and multiple doses of the selected medicament are released andinhaled.

[0049] The term “amount” as used herein refers to a quantity or to aconcentration as appropriate to the context. The amount of a drug thatconstitutes a therapeutically effective amount varies according tofactors such as the potency of the particular biotherapeutic medicament,the route of administration of the formulation, and the mechanicalsystem used to administer the formulation. A therapeutically effectiveamount of a particular drug can be selected by those of ordinary skillin the art with due consideration of such factors. Generally atherapeutically effective amount of biotherapeutic medicament will befrom about 0.00001 parts by weight to about 5 parts by weight based on100 parts by weight of the fluid or propellant selected.

[0050] A suitable fluid includes air, a hydrocarbon such as n-butane,propane, isopentane, etc. or a propellant. A suitable propellant is anyfluorocarbon, e.g. a 1-6 hydrogen containing flurocarbon (such asCHF₂CHF₂, CF₃CH₂F, CH₂F₂CH₃ and CF₃CHFCF₃), a perfluorocarbon, e.g. a1-4 carbon perfluorocarbon, (such as CF₃CF₃, CF₃CF₂CF₃); or any mixtureof the foregoing, having a sufficient vapor pressure to render themeffective as propellants. Some typical suitable propellants includeconventional chlorofluorocarbon (CFC) propellants such as propellants11, 12 and 114 or a mixture thereof. Non-CFC propellants such as1,1,1,2-tetrafluoroethane (Propellant 134a),1,1,1,2,3,3,3-heptafluoropropane (Propellant 227) or a mixture thereofare preferred. The fluid or propellant is preferably present in anamount sufficient to propel a plurality of selected doses of drug froman aerosol canister when such is employed.

[0051] A suitable first stabilizer is selected. A suitable firststabilizer includes (1) an amino acid selected from the group consistingof (a) a monoamino carboxylic acid of the formula, H₂N—R—COOH(I), (b) amonoamino dicarboxylic acid of the formula, H₂N—R(COOH)₂ (II) and (c) adiamino monocarboxylic acid of the formula (H₂N)₂—R COOH (III), where Ris a straight or branched alkyl radical of from 1 to 22 carbon atoms,which can be mono or poly-substituted with sulfide (—S—), oxide (—O—),hydroxyl (—OH), amide (—NH), sulfate (—SO4); aryl of the formula

[0052] where X is hydrogen, halogen alkyl of 1 to 6 carbon atoms, alkoxyof 1 to 6 carbon atoms, hydroxy and nitro; and heterocyclic, selectedfrom the groups consisting of thienyl, furyl, pyranyl, imidazolyl,pyrrolyl, thizolyl, oxazolyl, pyridyl, and pyrimidinyl; (2) a derivativeof the amino acid selected from (a) an acid addition salt of the aminogroup, as well as (c) an ester of the carboxylic acid group obtainedfrom aliphatic straight or branched chain alcohols of from 1 to 6 carbonatoms, (3) an ether of any of the foregoing; (4) a hydrate orsemi-hydrate of any of the foregoing and (5) a mixture of the amino acidand the derivative of the amino acid.

[0053] Suitable amino acids of the inventive formula include glycine,alanine, valine, leucine, isoleucine, leucylalanine, methionine,threonine, isovaline, phenylalanine, tyrosine, serine, cysteine,N-acetyl-L-cysteine, histidine, tryptophan, proline, and hydroxyproline,e.g. trans-4-hydroxy proline. Compounds of the formula II includeaspartic acid, and glutamic acid, compounds of the formula (III) includearginine, glutamine, lysine, hydroxylysine, omithine, asparagine, andcitrulline.

[0054] A fluid or aerosol formulation preferably comprises theprotective colloid stabilizer in an amount effective to stabilize theformulation relative to an identical formulation not containing thestabilizer, such that the drug does not settle, cream or flocculateafter agitation so quickly as to prevent reproducible dosing of thedrug. Reproducible dosing can be achieved if the formulation retains asubstantially uniform drug concentration for about 15 seconds to about15 minutes after agitation.

[0055] For optimal functional and therapeutic performance of the aerosolformulation, as an aerosol suspension, the stabilizer e.g., amino acidstabilizer, is present either as a coarse carrier (e.g., 20-90 μm) or asa finely micronized powder, ≦10 μm in diameter. In either case,reproducible drug dosimetry is obtained without the need to qualify theinspiratory maneuver of the patient. Accordingly, excellent doseuniformity is obtained at tidal flows of up to 2 liters, or atinspiratory flow rates of as low as 15 liters per minute to about 90liters per minute.

[0056] Alternatively, a second suitable stabilizer is selected. A secondsuitable stabilizer is a “water addition.” As used herein, a “wateraddition” is an amount of water which (1) is added, either initiallywith other components of the described aerosol formulation, e.g.medicament associated with the polymeric construct as part thereof orencapsulated therein, and fluid carrier, or after the other components,e.g. medicament, fluid carrier are combined and processed, (2) is inaddition to the water which is always present and which develops duringprocessing and/or storage of the aerosol formulation, i.e. “developed”or “nascent” formulation water, and (3) is present in an amount whichfurther stabilizes a medicinal aerosol formulation, e.g. rosiglitazonemaleate, having nascent formulation water.

[0057] An aerosol formulation preferably comprises the water addition inan amount effective to more effectively stabilize the formulationrelative to an identical formulation not containing the water addition,i.e. containing only nascent formulation water, such that the drug e.g.,an insulin containing construct, does not settle, cream or flocculateafter agitation so quickly as to prevent reproducible dosing of thedrug. Reproducible dosing can be achieved if the formulation retains asubstantially uniform drug concentration for about fifteen seconds toabout five minutes after agitation.

[0058] It is of course understood, that the first stabilizer can becombined with the second stabilizer in an approximate combinedconcentration, e.g. typically to 300 ppm to 2000 ppm, to achieve thedesired stability of the resultant formulation.

[0059] The particular amount of stabilizer e.g. amino acid stabilizeralone or water of addition stabilizer alone or both stabilizerscombined, that constitutes an effective amount is dependent upon theparticular stabilizer, the particular propellant, and on the particulardrug used in the formulation. It is therefore not practical to enumeratespecific effective amounts for use with specific formulations of theinvention, but such amounts can readily be determined by those skilledin the art with due consideration of the factors set forth above.Generally, however, the stabilizer can be present in a formulation in anamount from about 0.001 parts per million to about 200,000 parts permillion, more preferably about 1 part per million to about 10,000 partsper million, most preferably from about 10 parts per million to about5,000 parts per million of the total formulation.

[0060] It has surprisingly been found that the formulation of theinvention, e.g. suspension or solution is stable without the necessityof employing a cosolvent, such as ethanol, or surfactants. However,further components, such as conventional lubricants or surfactants,co-solvents, ethanol, etc., can also be present in an aerosolformulation of the invention in suitable amounts readily determined bythose skilled in the art. In this regard, reference is made to U.S. Pat.No. 5,225,183, which is incorporated by reference hereinto in itsentirety.

[0061] Generally the formulations of the invention can be prepared bycombining, matrixing, or encapsulating (i) the biotherapeutic medicamentor drug with a sufficient amount of the modulating polymer in an amountsufficient to provide a plurality of therapeutically effective doses ofthe biotherapeutic; (ii) if necessary, combining or adding anappropriate suspension stabilizer in an amount effective to stabilizeeach of the formulations; and (iii) dispersing the matrixed orencapsulated and stabilized biotherapeutic medicament in an appropriatefluid or propellant in an amount sufficient to propel a plurality ofdoses, e.g. from an aerosol canister.

[0062] A sufficient amount of the modulating polymer, e.g. an alginate,is dependent upon the desired rate of release of the medicament itself.Typically, for a release of about 2 to about 6 hours, the concentrationof the polymeric material ranges from about 10 ppm to about 100,000 ppmto effect a desired release profile e.g. about 15 minutes to about 12hours.

[0063] Particles of the selected polysaccharide polymer system may beprepared using solutions or emulsion preparations of the polymer andactive pharmaceutical ingredient which may subsequently be dried eitherby the use of an antisolvent such as carbon dioxide, nitrogen, or anyother appropriate antisolvent, or by solvent evaporation, spray drying,solvent extraction, phase separation, coacervation, interfacialpolymerization, and other methods well known to those of ordinary skillin the art. Polysaccharide polymeric particles may be made also usingmicroencapsulation, by nanoparticle technology, by coating methods suchas spray congealing, by supercritical fluid technology, or by micellarsolubilization where various techniques known to those skilled in theart may be used. These methods are described in the following non-exhaustive list of references which are incorporated hereinto byreference:

[0064] (1) Doubrow, M., Ed., “Microcapsules and Nanoparticles inMedicine and Pharmacy,” CRC Press, Boca Raton, 1992.

[0065] (2) Benita et al., J. Pharm. Sci. 73, 1721-1724 (1984);

[0066] (3) Cook et al., U.S. Pat. No. 4,044,126;

[0067] (4) Cook et al., U.S. Pat. No. 4,363,923;

[0068] (5) Cook et al., U.S. Pat. No. 4,414,209;

[0069] (6) Ecanow, U.S. Pat. No. 4,963,367;

[0070] (7) Hallworth et al., U.S. Pat. No. 4,206,758;

[0071] (8) Hallworth et al., U.S. Pat. No. 4,353,365;

[0072] (9) Lindsay, U.S. Pat. No. 5,169,433;

[0073] (10) Makiej, Jr., U.S. Pat. No. 5,002,048;

[0074] (11) Mathiowitz and Langer, J. Controlled Release 5,13-22 (1987);

[0075] (12) Mathiowitz et al., Reactive Polymers 6, 275-283 (1987);

[0076] (13) Mathiowitz et al., J. Appl. Polymer Sci. 35, 755-774 (1988);

[0077] (14) Mathiowitz et al., Scanning Microscopy 4: 329-340 (1990);

[0078] (15) Mathiowitz et al., J. Appl. Polymer Sci. 45, 125-134 (1992);

[0079] (16) Martin, U.S. Pat. No. 4,892,232;

[0080] (17) Newell et al., U.S. Pat. No. 4,811,73 1;

[0081] (18) Newell et al., U.S. Pat. No. 4,627,432;

[0082] (19) Ray, U.S. Pat. No. 4,800,903;

[0083] (20) Riccio, U.S. Pat. No. 3,856,185;

[0084] (21) Ronge, U.S. Pat. No. 5,056,511;

[0085] (22) Sievers et al., U.S. Pat. No. 4,970,093;

[0086] (23) Smith, U.S. Pat. No. 4,582,731;

[0087] (24) Whitsett, U.S. Pat. No. 5,013,720; and

[0088] (25) McNab, U.S. Pat. No., 5,044,523.

[0089] (26) Hanna and York, World Intellectual Property OrganizationPatent Number WO9959710A1

[0090] (27) Hanna, et al., World Intellectual Property OrganizationPatent Number W09944733A1

[0091] (28) Hanna and York, World Intellectual Property OrganizationPatent Number WO9836825A1

[0092] The modulated release particles of the invention can be deliveredto the respiratory tract and/or lung of the patient to be treated, e.g.a human being or other animal, by oral inhalation in order to effectbronchodilation or in order to treat a condition susceptible oftreatment by inhalation, e.g., asthma, chronic obstructive pulmonarydisease.

[0093] The modulated release particles of the invention can also bedelivered to the lung in order for the biotherapeutic agent to bedelivered at measured rates to the systemic circulation for treatment ofdiseases elsewhere in the body, e.g., diabetes, hormone replacement,cancer, erythropoiesis, infection, or for immune protection such asachievable with vaccines.

[0094] The modulated release particles of the invention can also bedelivered by nasal inhalation in order to treat, for example, allergicrhinitis, rhinitis, (local) or diabetes (systemic), or they can bedelivered via topical (e.g., buccal) administration in order to treat,e.g., angina or local infection.

[0095] Depending upon the concentration of the polymer, e.g. alginate,drug release rates range from about 5 minutes to several hours. Examplesof the release profile, and the corresponding glycemic control relativeto an intravenous administration in New Zealand rabbits is given inFIGS. 1 and 2.

We claim:
 1. A modulated release aerosol formulation, which comprises,a. a polymeric construct comprising a polysaccharide polymer having aselected medicament associated therewith; and b. a fluid carrier forcarrying and delivering said construct; and c. a stabilizer selectedfrom the group consisting of a first stabilizer, a second stabilizer anda mixture of the foregoing stabilizer.
 2. A modulated release aerosolformulation, which comprises, a. a selected particulate medicament; b.particles of a polysaccharide polymer having said medicament associatedtherewith c. a stabilizer selected from the group consisting of a firststabilizer, a second stabilizer and a mixture of the foregoingstabilizers combined with said particles; and d. a fluid carrier forcarrying and transporting said combined particles of copolymer.
 3. Theformulation as defined in claim 1 wherein said stabilizer comprises awater of addition.
 4. The formulation as defined in claim 1 wherein saidstabilizer is said first stabilizer selected from the group consistingof an amino acid selected from the group consisting of (a) a monoaminocarboxylic acid of the formula, H₂N—R—COOH(I), (b) a monoaminodicarboxylic acid of the formula, H₂N—R(COOH)₂ (II) and (c) a diaminomonocarboxylic acid of the formula (H₂N)₂—R COOH (III), where R is astraight or branched alkyl radical of from 1 to 22 carbon atoms, whichcan be mono or poly-substituted with sulfide (—S—), oxide (—O—),hydroxyl (—OH), amide (—NH), sulfate (—SO4); aryl of the formula

where X is hydrogen, halogen, alkyl of 1 to 6 carbon atoms, alkoxy of 1to 6 carbon atoms, hydroxy and nitro; and heterocyclic selected from thegroups consisting of thienyl, furyl, pyranyl, imidazolyl, pyrrolyl,thizolyl, oxazolyl, pyridyl, and pyrimidinyl; (2) a derivative of theamino acid selected from (a) an acid addition salt of the amino group,(b) an amide of the carboxylic acid group, (c) an ester of thecarboxylic acid group obtained from aliphatic straight or branched chainalcohols of from 1 to 6 carbon atoms, (3) an ether of any of theforegoing; (4) a hydrate or semi-hydrate of any of the foregoing and (5)a mixture of the amino acid and the derivative of the amino acid.
 5. Theformulation as defined in claim 1 wherein said first stabilizer is anamino acid selected from the group consisting of the twenty existingamino acids.
 6. The formulation as defined in claim 1 wherein saidpolymer is selected from the group consisting of alginic acid or apharmaceutically acceptable salt thereof; guar gum; gum karaya; gumBenjamin, plantago ovata gum; agar; carrageenan; cellulose; galaturonicacid or a mixture of any of the foregoing polymers.
 7. The formulationas defined in claim 1 wherein said medicament comprises a protein orpeptide medicament having a molecular size ranging from about 1K Daltonto about 150 K Daltons.
 8. The formulation as defined in claim 7 whereinsaid medicament is selected from the group consisting of an insulin, aninsulin analog, an amylin, an immunodilating protein, an interleukin, aninteferon, an erythropoietan, a heparin, a thrombolytic, an antitrypsin,an antiprotease, a hormone, a growth factor, an enzyme, a nucleic acid,an immunoglobulin, an antibiotic, an antiinfective, a calcitonin, ahematopoietic factor, a vaccine, a vasoactive peptide, an antisenseagent, an oligonucleotide, DNase, a cyclosporin, ribavirin or a mixtureof any of the foregoing medicaments.
 9. The formulation as defined inclaim 7 wherein said medicament is selected from the group consisting ofan insulin, an insulin analog, an amylin, glucagon, LH-RH, deltirex,leuprolide, gosorelin, nafarelin, octreotide, somatostatin, acalcitonin, porathyroid hormone, TRH, growth hormone-releasing hormone,G-CSF, G-SF, a cytokine, rhDNAse, a heparin, an oligoneucleotide,ribavarin, glucagon, acetohexamide, chlorpropamide, tolazemide,tolbutamide, glipizide, glyburide, glucophage, phentolamine, tumorneurosis factor (TNF), nerve growth factor (NGF), macrophage-colonystimulating factor (M-CSF), heparinase, bone morphogenic protein (BMP),hANP, glucagon-like peptide (GLP-1), renin, bradykinin, a bacitracin, apolymyxin, a colistin, tyrocidine, a gramicidin, a monoclonal antibody,a vaccine or a mixture of any of the foregoing medicaments.
 10. Theformulation as defined in claim 6 wherein said polymer is present in anamount of about 0.000001 to about 10 percent by weight of the totlaweight of the formulation.
 11. The formulation as defined in claim 1wherein said fluid carrier is selected from the group of propellantsconsisting of 1,1,1,2-tetrafluoethane, 1,1,1,2,3,3,3-heptafluoropropaneor a mixture thereof.
 12. The formulation as defined in claim 1 whereinsaid fluid carrier is a compressed gas selected from the groupconsisting of air, carbon dioxide, nitrogen and a mixture of any of theforegoing compressed gases.
 13. The formulation as defined in claim 1wherein said fluid carrier is a hydrocarbon selected from the groupconsisting of n-butane, propane, isopentene and a mixture of any of theforegoing hydrocarbons.
 14. The formulation as defined in claim 1wherein said fluid carrier is a chlorofluorocarbon selected from thegroup consisting of propellant 11, propellant 12, propellant 114 and amixture of any of the foregoing propellants.
 15. The formulation asdefined in claim 1 which further comprises a cosolvent.
 16. Theformulation as defined in claim 15 wherein said cosolvent comprisesethanol.
 17. The formulation as defined in claim 1 wherein saidstabilizer is present in an amount ranging from about 300 parts byweight to about 2000 parts by weight to the total weight of theformulation.
 18. A method of preparing a stable medicinal aerosolformulation according to claim 1, which comprises, a. combining saidselected medicament with said polymer to form said polymeric construct,wherein said medicament is associated with said polymer in an amountsufficient to provide a plurality of therapeutically effective doses; b.combining said stabilizer with said construct to form a stabilizercombined construct; c. combining said construct with a fluid carrier,where said carrier is present in an amount sufficient to transport anddeliver a plurality of said therapeutically effective doses, to form amixture; and d. dispersing said mixture.
 19. A method of treating in ahuman being or another animal a condition capable of treatment by oralor nasal inhalation, which comprises; administering a formulationaccording to claim 1 to said human being or animal by oral or nasalinhalation.
 20. A formulation according to claim 1 in an aerosolcanister equipped with a metered dose valve.
 21. A metered dose inhalercontaining a medicinal formulation, the formulation comprising a. aparticulate medicament in a therapeutically effective amount; b. apolysaccharide polymer with which said particulate medicament isassociated to form a particulate polymeric construct; c. a fluid carrierfor containing and transporting said particulate polymeric construct;and d. a suitable stabilizer selected from the group consisting of afirst stabilizer, a second stabilizer and a mixture of the foregoingstabilizers present in an amount sufficient to stabilize the formulationto prevent settling, creaming or flocculation thereof for a timesufficient to allow reproducible dosing of said associated medicamentafter agitation of the formulation.
 22. The inhaler as defined in claim21 wherein said stabilizer is selected from the group consisting of thetwenty existing amino acids, any mixture thereof and any derivative ofthe foregoing.
 23. The inhaler as defined in claim 22 wherein saidstabilizer is selected from the group consisting of (1) a di-peptideselected from a salt and an ester of oxidized and unoxidizedL-cysteinylglycine, gamma-L-glutamyl-L-cysteine,N-acetyl-L-cystein-glycine; (2) a conjugated, unconjugated or polymericform of L-Gly-L-glu and L-Val-L-Thr; (3) L-aspartyl-L-phenylamine; (4) amuramyl dipeptide; (5) a nutrient selected from L-tyrosyl-L-tyrosine,L-alanyl-L-tyrosine, L-arginyl-L-tyrosine, L-tyrosyl-L-arginin,N-Clz-L-Liu-OCH and salts and esters of the foregoing; (6)glycyl-glycine; (7) N-acetyl-L-aspartate-L-glytamate (NAAG); (8) atripeptide selected from an oxidized and an unoxidized form ofgamma-L-glutamyl-L-cysteine glycine or a muramyl tripeptide and (9) amixture of any of the foregoing stabilizers.
 24. The inhaler as definedin claim 21 wherein said stabilizer is the second stabilizer whichcomprises a water of addition.
 25. The formulation as defined in claim 5wherein said stabilizer is selected from the group consisting of (1) adi-peptide selected from a salt and an ester of oxidized and unoxidizedL-cysteinylglycine, gamma-L-glutamyl-L-cysteine,N-acetyl-L-cystein-glycine; (2) a conjugated, unconjugated or polymericform of L-Gly-L-glu and L-Val-L-Thr; (3) L-aspartyl-L-phenylamine; (4) amuramyl dipeptide; (5) a nutrient selected from L-tyrosyl-L-tyrosine,L-alanyl-L-tyrosine, L-arginyl-L-tyrosine, L-tyrosyl-L-arginin,N-Clz-L-Liu-OCH and salts and esters of the foregoing; (6)glycyl-glycine; (7) N-acetyl-L-aspartate-L-glytamate (NAAG); (8) atripeptide selected from an oxidized and an unoxidized form ofgamma-L-glutamyl-L-cysteine glycine or a muramyl tripeptide and (9) amixture of any of the foregoing stabilizers.